Immune responses against hepatitis C virus genotype 3a virus-like particles in mice: A novel VLP prime-adenovirus boost strategy

Chronic hepatitis C virus (HCV) infection represents a major health threat to global population. In India, approximately 15–20% of cases of chronic liver diseases are caused by HCV infection. Although, new drug treatments hold great promise for HCV eradication in infected individuals, the treatments are highly expensive. A vaccine for preventing or treating HCV infection would be of great value, particularly in developing countries. Several preclinical trials of virus-like particle (VLP) based vaccine strategies are in progress throughout the world. Previously, using baculovirus based system, we have reported the production of hepatitis C virus-like particles (HCV-LPs) encoding structural proteins for genotype 3a, which is prevalent in India. In the present study, we have generated HCV-LPs using adenovirus based system and tried different immunization strategies by using combinations of both kinds of HCV-LPs with other genotype 3a-based immunogens. HCV-LPs and peptides based ELISAs were used to evaluate antibody responses generated by these combinations. Cell-mediated immune responses were measured by using T-cell proliferation assay and intracellular cytokine staining. We observed that administration of recombinant adenoviruses expressing HCV structural proteins as final booster enhances both antibody as well as T-cell responses. Additionally, reduction of binding of VLP and JFH1 virus to human hepatocellular carcinoma cells demonstrated the presence of neutralizing antibodies in immunized sera. Taken together, our results suggest that the combined regimen of VLP followed by recombinant adenovirus could more effectively inhibit HCV infection, endorsing the novel vaccine strategy.     

河南省1例柬埔寨输入登革热病例的病原分子分型与全基因序列分析

目的鉴定河南省1例来自柬埔寨的登革热输入性病例的登革病毒(dengue virus,DENV)血清型和基因型及其序列特征和传播来源。方法患者血液标本来源于2019年国家疾病监测系统网络直报的登革热疑似病例。样品经快速检测DENV NS1抗原和IgM/IgG抗体,再提取血清中核酸,应用荧光RT-PCR法进行DENV血清型鉴定,同时用Vero和BHK-21细胞对血清标本进行病毒分离培养,阳性培养物扩增全基因组序列,进行序列系统进化分析。结果该病例实验室确诊为DENV 1型感染,并从血清标本中分离到病毒株,测序后拼接成全长10670 nt的全基因组序列,经系统进化分析,该病毒株属于DENV 1型基因Ⅰ型,与东南亚流行株具有较高同源性和较近亲缘关系。结论2019年河南省来自柬埔寨的输入性病例的病原体为DENV 1型基因I型,此为近年来东南亚输入我国的常见血清型和基因型。     

直接抗病毒药物治疗丙型肝炎合并透析患者的临床应用

丙型肝炎病毒（HCV）感染在慢性肾功能不全患者中较常见，尤其是在进行透析的患者中，HCV感染较普通患者的风险极大增加，同时还会导致肝细胞癌及肝硬化的发病率明显升高。近年来直接抗病毒药物（DAAs）在慢性丙型肝炎的治疗中取得了较好的疗效及安全性，本文通过总结DAAs在丙型肝炎合并透析患者中的应用进展，发现对于基因1~6型HCV合并透析患者，推荐使用G/P方案；由于索磷布韦（SOF）经肾脏代谢，在重度肾损害人群中的血药浓度较高，因此SOF组合方案并不推荐用于丙型肝炎合并透析患者的治疗。     

Expression of TRIM22 mRNA in chronic hepatitis C patients treated with direct-acting antiviral drugs

Hepatitis C is a global public health problem, and Pakistan is the second largest country in the globe with highest prevalence rate of hepatitis C virus (HCV). Until 2014, pegylated interferon (PEG-IFN) plus ribavirin (RBV) has been the standard therapy for HCV, however, owing to its adverse side effects and very low sustained virologic response (SVR) rates therapeutics trend is shifted toward direct-acting antivirals. Tripartite motif containing 22 (TRIM22) is a dynamic antiviral protein that can inhibit multiple viruses in vivo. Expression of TRIM22 mRNA has been linked to outcome of PEG-IFN and ribavirin therapy, where its higher expression leads to rapid virus clearance. However, in terms of therapy with direct-acting antiviral (DAA) or double DAA, impact of TRIM22 expression is largely unknown. These new drugs show more than 90% of SVR rates and lesser side effects and have proven to be better than IFN therapy. Endogenous IFN system suppresses various pathogens through the induction of antiviral effectors termed as interferon-stimulating genes (ISGs). We have studied the expression levels of one of these antiviral effectors, TRIM22 in response to sofosbuvir (SOF) and daclatasvir (DAC) in combination with RBV, using quantitative PCR in the peripheral blood mononuclear cells (PBMCs) of HCV-infected patients. We have observed sustained virus clearance in more than 90% of patients treated with DAA and double DAA and have seen the expression of TRIM22 to be higher in patients who attained SVR as compared to the untreated patients. We have also observed downregulation of TRIM22 in patients who failed to attain rapid virus clearance, and upregulation in those who achieved rapid clearance of virus. Genetic factors that determine the lower TRIM22 expression in these patients are needed to be explored that may also play a role in lower response to anti-HCV therapy. Endogenous IFN system and effects of antiviral proteins in response to DAA therapy is needed to be studied in order to better understand the host response toward these drugs to make them more effective.     

桑根酮C对博莱霉素诱导小鼠肺纤维化改善作用及机制研究

目的观察桑根酮C对博莱霉素诱导的小鼠肺纤维化的影响,并探讨其可能的作用机制。方法 C57BL/6小鼠随机分为4组,即假手术组、模型组和桑根酮C(100、50 mg/kg)组,每组20只。假手术组小鼠气管内注射生理盐水,模型组小鼠气管内注射博莱霉素诱导肺纤维化模型。术后第4天开始给药,连续给药28d后检测小鼠呼吸功能;检测肺内羟脯氨酸含量;HE染色观察肺内炎症表现;Masson染色观察肺内胶原活性;免疫组化法检测肺内转化生长因子-β1(TGF-β1)蛋白表达量;免疫蛋白印迹法检测肺内α平滑肌肌动蛋白(α-SMA)、核转录因子-κB p65(NF-κB p65)、磷酸化的核转录因子-κB p65(p-NF-κB p65)、Ⅰ型胶原和Ⅲ型胶原表达量。结果与模型组比较,桑根酮C能够改善经博莱霉素诱导形成肺纤维化后小鼠的呼吸功能,能够明显降低肺内羟脯氨酸含量,明显减轻肺内炎症和胶原沉积,肺内TGF-β1、α-SMA、NF-κBp65、p-NF-κBp65、Ⅰ型胶原和Ⅲ型胶原蛋白表达量明显降低。结论桑根酮C能够明显改善博莱霉素诱导小鼠肺纤维化,改善呼吸功能,其机制可能与抑制TGF-β1过表达和降低炎症转录因子NF-κB及磷酸化表达有关。     

基于机器学习的脑胶质瘤多模态影像分析

脑部胶质瘤是临床中常见的一种原发性脑肿瘤，具有复发率高、死亡率高以及治愈率低的特点。常规临床诊断主要依靠计算机断层扫描(CT)和磁共振成像(MRI)检查技术进行鉴别。随着成像技术和机器学习方法的不断发展，多模态影像智能分析技术已经逐步成为研究热点，在脑胶质瘤的病灶分割测量、肿瘤分级、预后生存周期预测和基因型辨别等方面具有重要的应用前景。本文重点介绍基于机器学习和多模态影像在脑胶质瘤临床辅助诊断和预后评估中的应用进展。     

Endodontic treatment of a hypertaurodontic mandibular left second molar in a patient with many taurodonts combined with multiple pulp stones

Taurodontism is a rare embryologic anomaly of teeth, defined by an apical displacement of the furcation of roots and enlarged pulp chambers. Taurodontism has been classified as hypo‐, meso‐ or hypertaurodontism according to the severity of the anomaly. The aim of this case report was to illustrate a clinical case with multiple bilateral taurodonts and to describe the endodontic management of the hypertaurodontic mandibular left second molar with a C‐shaped canal and extensive dental pulp calcifications. A healthy 20‐year‐old male patient was referred for the endodontic treatment of his lower left second molar. Cone beam computed tomography revealed a C‐shaped root canal configuration and several dental pulp calcifications in this tooth. The endodontic treatment was performed in two appointments under an operating dental microscope. A panoramic radiograph, made during the 18 months follow‐up appointment, revealed nine other taurodontic molars, most of them associated with dental pulp calcifications.